2.Effect of melatonin on the proliferation, apoptosis, and expression of bcl-2 in oxidized low-density lipoprotein-induced endothelial progenitor cells.
Xiu-li LI ; Xiu-mei XIE ; Xiao-bin CHEN ; Jin HE ; Ye-qing FANG
Journal of Central South University(Medical Sciences) 2007;32(5):862-867
OBJECTIVE:
To explore the effect of melatonin(Mel) on the proliferation, apoptosis and expression of bcl-2 in oxidized low-density lipoprotein(ox-LDL)-induced endothelial progenitor cells (EPC) from human umbilical cord blood in vitro.
METHODS:
Total mononuclear cells were isolated from human umbilical cord blood in vitro by Ficoll density gradient centrifugation, and the cells were plated on fibronectin-coated culture dishes. After 7 days, the attached cells were divided into 7 groups: a control group (normal cells), 3 ox-LDL groups[the attached cells were incubated with different concentrations of ox-LDL(5,10,and 20mg/L) for 24 hours], and 3 Mel groups[the attached cells were incubated with different concentrations of Mel (0.5,1.0, and 2.0 mmol/L) respectively for 24 hours before incubation with 10 mg/L ox-LDL]. EPC was identified by examining the expression of CD34, vascular endothelial growth factor receptor-2(VEGFR-2) and CD133 under a laser scanning confocal microscope. We used 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay to detect the effect of Mel and ox-LDL on the multiplication ability of EPC. Flow cytometry was used to detect the apoptosis. The expressions of Bcl-2 mRNA and protein were detected respectively by RT-PCR and immunohistochemistry technology.
RESULTS:
After being exposed to the ox-LDL, the proliferation of EPC in the 3 ox-LDL groups was lower, and the apoptosis rate was higher than that in the control group in a dose-dependent manner (P<0.01); Mel was added at different concentrations before the ox-LDL incubation, and the cells in the 3 Mel groups showed higher proliferation and lower apoptosis rate than those of the 3 ox-LDL groups (P<0.01). Expression of Bcl-2 mRNA and protein of EPC in the 3 Mel groups was higher than that in the 3 ox-LDL groups (P<0.01).
CONCLUSION
Ox-LDL can inhibit the proliferation of EPC and promote the apoptosis of the cells by down-regulating the bcl-2 expression. Mel can inhibit these effects of ox-LDL.
Apoptosis
;
drug effects
;
Cell Proliferation
;
drug effects
;
Cells, Cultured
;
Human Umbilical Vein Endothelial Cells
;
cytology
;
drug effects
;
Humans
;
Lipoproteins, LDL
;
adverse effects
;
Melatonin
;
pharmacology
;
Proto-Oncogene Proteins c-bcl-2
;
metabolism
;
Stem Cells
;
cytology
;
drug effects
3.Change of endothelial progenitor cells from peripheral blood in patients with coronary heart diseases before and after percutaneous coronary intervention.
Xiao-bin CHEN ; Xiao-qun PU ; Xiu-mei XIE ; Ye-qing FANG ; Jin HE ; Xiu-li LI
Journal of Central South University(Medical Sciences) 2008;33(5):432-437
OBJECTIVE:
To investigate the culture of endothelial progenitor cells (EPCs) from peripheral blood in patients with coronary heart diseases (CHD) before and after percutaneous coronary intervention (PCI), and to observe the cells shape and determine the cell number and proliferation activity.
METHODS:
Ninety-five patients were divided into a CHD group(n=65) and a control group (n=30). The mononuclear cells were isolated from peripheral blood of patients with CHD before, right after and 4 days after PCI by Ficoll-density centrifugation. The isolated cells were cultured in RPMI1640 medium supplemented with VEGF165 and bFGF.EPCs were characterized as adherent cells of double positive for DiL-acLDL uptake and FITC-UEA-I binding by direct fluorescent staining under a fluorescence microscope. The EPCs specific surface mark CD34 and KDR were assessed by fluorescence activated cell sorter analysis. The cell shapes were analysed and the number of colony-forming units(CFU) was counted by phase-contrast microscope.
RESULTS:
The number of EPCs reduced in patients with CHD before the PCI, but the cell number was significantly increased in patients with CHD after the PCI, and the number reduced in patients with CHD 4 days after the PCI. How-ever, the number of CFUs did not change in patients before and after the PCI.
CONCLUSION
PCI can increase endothelial progenitor cells in patients after the PCI; but 4 days after the PCI, this increase will not exist.
Aged
;
Aged, 80 and over
;
Angioplasty, Balloon, Coronary
;
Cell Adhesion
;
Cell Count
;
Cell Movement
;
Cells, Cultured
;
Coronary Disease
;
blood
;
therapy
;
Endothelial Cells
;
pathology
;
Female
;
Humans
;
Male
;
Middle Aged
;
Stem Cells
;
pathology
4.Study on the status of pollution by Legionella species in hot spring vacation center and the related factors.
Lian-Xiu WANG ; Ge-Bin YAN ; Li-Na GE ; Gui-Fang QIAO
Chinese Journal of Epidemiology 2006;27(11):943-946
OBJECTIVETo study the pollution status of Legionella species in hot spring vacation center and the related factors.
METHODSField surveys were performed in four big hot spring vacation centers of Changping district. Uniform questionnaires was used and colony count was made together with the isolation of Legionella species from hot spring water based on mip gene typing.
RESULTS47 isolates of Legionella pneumophila (Lp) from 87 samples showed 4 serotypes as Lp1, Lp6, Lp12, Lp5 with percent of 57.45%, 21.28%, 14.89%, 6.38% respectively. The hot spring centers controlled the temperature of recycled water between 34-47 degrees C by hot water heating and filtrating system. All the isolates were cultured from the hot water with temperature between 34-44 degrees C: 56.75% (21/37) in high temperature (40-47 degrees C) and 61.90% (26/42) in low temperature (34-39.9 degrees C). There were no statistically significant difference between the high and the low temperature samples (P > 0.05). In the four hot spring vacation centers, the pH value was under control at 6.4-7.3 and the ambient temperature was under control between 26-28 degrees C. The humidity was controlled between 56% -69% relative humidity, which were the best growing conditions for the Legionella species. Disinfectors as chlorine deviratives was used in the four hot spring vacation centers. Though the concentration of chlorine in the water was 0.3-0.5 mg/L, 14.29%-48.00% of the samples were still positive of having Legionella species.
CONCLUSIONThe pollution of Legionella species was considered to be quite serious in the four hot spring vacation centers and the predominant serotype was Lp1. The pH and temperature of the hot spring water, ambient temperature and humidity and the way of heating up the water were the best conditions for the growth of Legionella species in these centers. Because of the high temperature of the hot spring water, chlorine of the disinfector volatilized quickly, affecting the effect of disinfection. The result revealed that water temperature achieving 44 degrees C could have had the effect of prevention.
China ; Disinfection ; Environmental Monitoring ; Hot Springs ; microbiology ; Legionella ; growth & development ; isolation & purification ; Temperature ; Travel ; Water Microbiology
5.Studies of betuionic acid on cell cycle and related protein expressions on mice of bearing H22 tumor cells.
Xiu-juan ZHANG ; Lei HAN ; Yu-bin JI ; Gui-zhen FANG
China Journal of Chinese Materia Medica 2008;33(14):1739-1743
OBJECTIVETo study the betulonic acid on the cell cycle and related protein expressions on mice of bearing H22 tumor cells.
METHODFlow cytometray was used to observe the changes of betulonic acid on the cell cycle and P53 of H22 tumor cells. Immunohistochemistry was determined the espressions of PI3K and AKT.
RESULTIncreasing the doses of betulonic acid, the number of H22 cells in S phase and G2 phase was increasing gradually, it can speculate that when the betulonic acid act on cells, the cells were blocked in S and G2 phase and inhibited the protein expressions of PI3K and AKT.
CONCLUSIONBetulonic acid may be up-regulate the activity of P53 and inhibite the expressions of PI3K and AKT, so that it inhibited the survival pathway of tumor cells.
Animals ; Betula ; chemistry ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Survival ; drug effects ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Female ; Flow Cytometry ; G2 Phase ; drug effects ; Gene Expression Regulation ; drug effects ; Immunohistochemistry ; Male ; Mice ; Neoplasm Transplantation ; Phosphatidylinositol 3-Kinases ; metabolism ; Proto-Oncogene Proteins c-akt ; metabolism ; Tumor Suppressor Protein p53 ; metabolism
6.Identification of rice seed varieties using neural network.
Zhao-yan LIU ; Fang CHENG ; Yi-bin YING ; Xiu-qin RAO
Journal of Zhejiang University. Science. B 2005;6(11):1095-1100
A digital image analysis algorithm based color and morphological features was developed to identify the six varieties (ey7954, syz3, xs11, xy5968, xy9308, z903) rice seeds which are widely planted in Zhejiang Province. Seven color and fourteen morphological features were used for discriminant analysis. Two hundred and forty kernels used as the training data set and sixty kernels as the test data set in the neural network used to identify rice seed varieties. When the model was tested on the test data set, the identification accuracies were 90.00%, 88.00%, 95.00%, 82.00%, 74.00%, 80.00% for ey7954, syz3, xs11, xy5968, xy9308, z903 respectively.
Algorithms
;
Artificial Intelligence
;
Cluster Analysis
;
Colorimetry
;
methods
;
Image Enhancement
;
methods
;
Image Interpretation, Computer-Assisted
;
methods
;
Information Storage and Retrieval
;
methods
;
Oryza
;
anatomy & histology
;
classification
;
Pattern Recognition, Automated
;
methods
;
Photography
;
methods
;
Reproducibility of Results
;
Seeds
;
anatomy & histology
;
classification
;
Sensitivity and Specificity
;
Species Specificity
7.Expression of angiopoietin-1 and its tyrosine kinase receptor Tie-2 in the airway of asthmatic rats.
Jun-Ying QIAO ; Bin LUAN ; Su-Ge HAN ; Xiu-Fang WANG
Chinese Journal of Contemporary Pediatrics 2008;10(5):642-646
OBJECTIVETo study the effect of dexamethasone on airway morphology and on the expression of angiopoietin-1 (Ang-1) and its tyrosine kinase receptor Tie-2 in the airway of asthmatic rats.
METHODSForty-five Sprague-Dawley rats were randomly divided into control, asthmatic, and dexamethasone-treated asthmatic groups. Asthma was induced by repeated sensitization and challenge with ovalbumin in the latter two groups. The dexamethasone intervention group received an intraperitonea injection of dexamethasone (2 mg/kg) before asthma challenge. Immunohistochemistry was used to measure the expression of Ang-1 and Tie-2 in the airway. Airway thickness was estimated by a computerized digital image analyzer.
RESULTSAirway thickness in the asthmatic group (33.9333+/-8.3791 micro m2/micro m) increased significantly compared with that in the control group (21.1333+/-2.7740 micro m2/micro m) (P<0.01). The dexamethasone intervention group also showed increased thickness of the airway (27.4000 +/- 4.6105 micro m2/micro m) compared with the control group (P<0.01), but the airway thickness in the dexamethasone intervention group was significantly reduced compared with that in the untreated asthmatic group (P<0.01). The expression of Ang-1 (103.9487+/-8.2914 vs 76.0320+/-3.7728; P<0.01) and Tie-2 (99.2307+/-8.1913 vs 75.3153+/-3.7321; P<0.01) in the airway increased significantly in the asthmatic group compared to controls. The expression of Ang-1 and Tie-2 in the airway of the dexamethasone intervention group (90.6180+/-5.2339 and 86.6633+/-3.7321, respectively) was statistically higher than that in the control group (P<0.01) but statistically lower than that in the untreated asthmatic group (P<0.01). Ang-1 and Tie-2 expression in the airway was positively correlated with the thickness of airway (r(Ang)-1=0.719r(Tie)-2=0.746P<0.01). There was also a positive correlation between Ang-1 and Tie-2 expression (r=0.742P<0.01).
CONCLUSIONSThe expression of Ang-1 and Tie-2 in the airway increased in asthmatic rats and was positively correlated with the thickness of the airway. Ang-1 and Tie-2 may participate in the process of airway remodeling in asthma. Dexamethasone can decrease the expression of Ang-1 and Tie-2 in the airway and relieve the changes of airway morphology.
Angiopoietin-1 ; analysis ; physiology ; Animals ; Asthma ; metabolism ; pathology ; Female ; Lung ; chemistry ; pathology ; Rats ; Rats, Sprague-Dawley ; Receptor, TIE-2 ; analysis ; physiology
8.A preliminary study on the genetic mode of aggressive periodontitis in Chinese Han nationality.
Xiu-yun REN ; Li XU ; Huan-xin MENG ; Rui-fang LU ; Zhi-bin CHEN ; Xiang-hui FENG
Chinese Journal of Stomatology 2012;47(2):75-80
OBJECTIVETo investigate the potential genetic mode of aggressive periodontitis (AgP) in Chinese Han nationality.
METHODSA total of 233 subjects from 73 nuclear families were recruited. All probands were diagnosed according to the criteria of AgP in 1999 classification of periodontal diseases. Ninety parents, 35 siblings and three grandparents and two offspring were examined based on full-mouth periodontal chartings (including parameter of probing depths, attachment loss, bleeding on probing at six sites per tooth) and full-mouth periapical radiographs. The genetic ratio was calculated and analyzed by the methods of Edwards and simple segregation.
RESULTSThe prevalence of AgP in probands' siblings was close to the square root of the prevalence of general population. The segregation ratio was 0.2419, which was close to the theoretical ratio for autosomal recessive inheritance. However, autosomal dominant inheritance could not be rejected in families whose parent(s) suffered from severe chronic periodontitis.
CONCLUSIONSThe genetic heterogeneity of AgP existed in Chinese Han nationality. The genetic mode was autosomal recessive inheritance in general, and autosomal dominant inheritance could not be excluded in families whose parent(s) suffered from severe chronical periodontitis. The results imply the genetic heterogeneity of AgP, and further demonstrate that AgP was a multifactorial disease with major genetic component in the disease etiology.
Aggressive Periodontitis ; epidemiology ; genetics ; Asian Continental Ancestry Group ; genetics ; Chronic Periodontitis ; epidemiology ; genetics ; Female ; Genes, Dominant ; Genes, Recessive ; Genetic Heterogeneity ; Humans ; Male ; Pedigree ; Prevalence ; Surveys and Questionnaires
9.Short- and long-term therapeutic effects of combination therapy with perindopril and irbesartan in a rat model of dilated cardiomyopathy.
Guo-Tian MA ; Xiu-Mei XIE ; Xiang-Hong WU ; Xiao-Bin CHEN ; Ye-Qing FANG ; Jin HE
Journal of Central South University(Medical Sciences) 2007;32(4):594-598
OBJECTIVE:
To evaluate the short-term, and long-term therapeutic effects of combination therapy with perindopril and irbesartan in a rat model of dilated cardiomyopathy (DCM).
METHODS:
Sprague-Dawley rats were administered adriamycin intraperitoneally to develop DCM. Grouping of rats: Group A contained normal rats, and Group B contained DCM rats. Both Group A and B were not given drug treatment. Group C and D contained DCM rats, however, Group C was administered perindopril 2mg/(kg x d) while Group D was administered perindopril 1mg/(kg x d) and irbesartan 25mg/(kg x d). Brain natriuretic peptide (BNP) was determined by enzyme linked immunosorbent assay; plasma potassium and creatinine were measured; the pathological lesions of cardiac muscle tissues were evaluated after HE staining; and the survival time of each rat during the intervention was recorded.
RESULTS:
After the three-week intervention, the plasma concentrations of BNP in Group D were lower than those in Group C (P<0.05). In each group, plasma concentrations of potassium and creatinine showed no significant differences between pre-intervention and post-intervention (P>0.05); pathological lesions of cardiac muscle tissues in both Group C and D were attenuated compared with those in Group B (P<0.01), but pathological lesions of cardiac muscle tissues showed no significant differences between Group C and Group D (P>0.05). Log-rank test showed that the life span of Group C was shorter than that of Group D (P<0.05); Cox regression analysis showed that both combination therapy and monotherapy with perindopril could prolong the survival time, but the effect of combination therapy was more obvious.
CONCLUSION
Combination therapy with perindopril and irbesartan in a rat model of DCM can more effectively improve the cardiac function and long-term prognosis than those monotherapy with perindopril. Both these two treatment plans can attenuate the pathological lesions of cardiac muscle tissues, without elevating the concentrations of plasma potassium and creatinine.
Animals
;
Biphenyl Compounds
;
therapeutic use
;
Cardiomyopathy, Dilated
;
chemically induced
;
drug therapy
;
Creatinine
;
blood
;
Doxorubicin
;
adverse effects
;
Drug Therapy, Combination
;
Irbesartan
;
Male
;
Myocardium
;
pathology
;
Natriuretic Peptide, Brain
;
metabolism
;
Perindopril
;
therapeutic use
;
Potassium
;
blood
;
Rats
;
Rats, Sprague-Dawley
;
Tetrazoles
;
therapeutic use
;
Treatment Outcome
10.Differentiation of 2 endothelial progenitor cells in vitro and inhibitory effect of asymmetric dimethylarginine on its proliferation.
Jin HE ; Xiu-mei XIE ; De-jian JIANG ; Ye-qing FANG ; Xiao-bin CHEN
Journal of Central South University(Medical Sciences) 2008;33(2):138-145
OBJECTIVE:
To investigate the Methods for culturing two types of endothelial progenitor cells (EPC) from human umbilical cord blood and study their differentiation traits and the depressant effect of asymmetric dimethylarginine (ADMA) on its proliferation.
METHODS:
Mononuclear cells were isolated from fresh cord blood by 6% hydroxyethyl starch(HES) and density gradient centrifugation.Isolated cells were cultured in the medium supplemented with vascular endothelial growth factors (VEGF) and basic fibroblast growth factors (bFGF). The growth characteristics and biological features of the cells were observed at different time points and identified by morphology,immunofluorescence staining,reverse transcription polymerase chain reaction (RT-PCR), and flow cytometry.Attached cells were incubated with different concentrations of ADMA (1,5, and 10 micromol/L) for 24,48, and 72 hours. Methylthiazoletetrazolium (MTT) assay and quantified colony forming units (CFUs) were used to assess the proliferation of endothelial progenitor cells.
RESULTS:
The attached cells were divided into 2 types:early EPC and late EPC. Early EPC changed from small sized round cells to spindle shaped cells and late EPC formed a typical cobblestone-like cells. Fluorescence microscopy showed that EPC were positive for both Dil-acLDL uptake and FITC-UEA-I binding.RT-PCR and FACS showed the difference of endothelial cell-specific,gene expression and changed AC133,CD34, and KDR among different times.Incubation of EPC with ADMA dose and time-dependently decreased the number and the proliferation of EPC.
CONCLUSION
There are 2 types of EPC from a source of human umbilical cord blood and ADMA may depress the EPC proliferation, providing a basis for further research.
Arginine
;
analogs & derivatives
;
pharmacology
;
Cell Differentiation
;
Cell Proliferation
;
drug effects
;
Cells, Cultured
;
Culture Media
;
Depression, Chemical
;
Endothelial Cells
;
cytology
;
Fetal Blood
;
cytology
;
Humans
;
Leukocytes, Mononuclear
;
cytology
;
Stem Cells
;
cytology